Quorum Sensing-Regulated Phenol-Soluble Modulins Limit Persister Cell Populations in Staphylococcus aureus.
Incomplete killing of bacterial pathogens by antibiotics is an underlying cause of treatment failure and accompanying complications. Among those avoiding chemotherapy are persisters being individual cells in a population that for extended periods of time survive high antibiotic concentrations proposedly by being in a quiescent state refractory to antibiotic killing. While investigating the human pathogen Staphylococcus aureus and the influence of growth phase on persister formation, we noted that spent supernatants of stationary phase cultures of S. aureus or S. epidermidis, but not of distantly related bacteria, significantly reduced the persister cell frequency upon ciprofloxacin challenge when added to exponentially growing and stationary phase S. aureus cells. Curiously, the persister reducing activity of S. aureus supernatants was also effective against persisters formed by either S. carnosus or Listeria monocytogenes. The persister reducing component, which resisted heat but not proteases and was produced in the late growth phase in an agr quorum-sensing dependent manner, was identified to be the phenol-soluble modulin (PSM) toxins. S. aureus express several PSMs, each with distinct cytolytic and antimicrobial properties; however, the persister reducing activity was specifically linked to synthesis of the PSMα family. Correspondingly, a high-persister phenotype of a PSMα mutant was observed upon fluoroquinolone or aminoglycoside challenge, demonstrating that the persister reducing activity of PSMs can be endogenously synthesized or extrinsically added. Given that PSMs have been associated with lytic activity against bacterial membranes we propose that PSM toxins increase the susceptibility of persister cells to killing by intracellularly acting antibiotics and that chronic and re-occurring infections with quorum sensing, agr negative mutants may be difficult to treat with antibiotics because of persister cells formed in the absence of PSM toxins.


Determination of dielectric properties of plaster blocks for sealing masonry using non-destructive frequency scanning methods.
With the increasing use of traditional and new models of wireless communication systems, the study and determination of the electrical characteristics of materials used in civil construction is an important topic to establish an understanding of how the radio frequency signal behaves inside built environments. This study presents an extensive process of characterization of electrical parameters of plaster blocks used in the construction of walls. Different from the literature where a prior estimation of the data occurs to enable sampling parameters to be obtained, this work proposes an innovative way of obtaining them entirely based on the analysis of the material through frequency measurements, which results in a greater level of precision of the data results. Analyses are made in the frequency range from 0.7 to 5.2 GHz, which has been used for several wireless communication standards. To carry out the electrical characterization of dielectric materials, a non-invasive methodology is proposed based on an innovative combination of the Nicolson-Ross-Weir Method (NRW) and the Ray Tracing Method. Through the proposed methodology and an extensive campaign of measurements using frequency scanning equipment, calculated and experimental data of Shielding Effectiveness (SE), complex relative electrical permittivity, loss tangent, attenuation coefficient and conductivity were obtained for a wide range of frequency, considering different samples of plaster blocks. The obtained results are compared to those available in the related literature, confirming the accuracy of the proposed analysis.


Nucleotide sequence and characterization of toxR: a gene involved in exotoxin A regulation in Pseudomonas aeruginosa.
We have previously reported the discovery and subsequent cloning of a regulatory gene, designated toxR, which appears to regulate the expression of the exotoxin A (ETA) structural gene toxA. Subsequent work by this laboratory has resulted in the subcloning of the toxR gene and its transfer to a high copy number plasmid (pGW28). Functional analysis of the toxR gene using a Tn5 insertion along with toxR deletions indicates that inactivation of toxR results in a dramatic reduction of ETA production. Nucleotide sequence analysis of pGW28 has revealed a 675 bp major open reading frame (225 codons) which could encode for a protein of 24,626 daltons. Using S1 nuclease mapping, the toxR RNA transcript has been shown to originate 20 bp upstream of the presumptive translation initiation codon. Experiments using a toxA specific probe have revealed the the toxR gene product appears to regulate the expression of ETA at the transcriptional level.


Potential risk of isoflavones: toxicological study of daidzein supplementation in piglets.
As a main component of soy isoflavones, daidzein is rich in soy-derived products, which are widely used as feed ingredients in farm animals. However, little research has been conducted on the side effects of dietary daidzein, especially in young animals. In this study, the safety of daidzein was evaluated. Results show that ingesting 400 mg/kg of dietary daidzein for 70 days is associated with a lower average daily weight gain (kilogram) (0.47 ± 0.03 vs 0.54 ± 0.04, P < 0.05) and a higher splenic damage index (1.00 ± 1.10 vs 0.00 ± 0.00, P < 0.05) in young pigs compared with control. Female pigs receiving 200 and 400 mg/kg daidzein showed reduced serum testosterone levels (ng/L) on days 35 and 70 compared with the control group (day 35, 246 ± 74 and 224 ± 20 vs 362 ± 48, P < 0.05; day 70, 252 ± 38 and 219 ± 77 vs 374 ± 38, P < 0.05). Daidzein residue (μg/kg) in pig livers increased (243 ± 80 vs 142 ± 47, P < 0.05, day 70). These results suggest that dietary supplements of 400 mg/kg of daidzein negatively affect the weight gain and splenic morphology of pigs.


Functional expression of the GABAB receptor in human airway smooth muscle.
gamma-Aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the mammalian central nervous system and exerts its actions via both ionotropic (GABA(A)/GABA(C)) and metabotropic (GABA(B)) receptors (R). In addition to their location on neurons, GABA and functional GABA(B) receptors have been detected in nonneuronal cells in peripheral tissue. Although the GABA(B)R has been shown to function as a prejunctional inhibitory receptor on parasympathetic nerves in the lung, the expression and functional coupling of GABA(B) receptors to G(i) in airway smooth muscle itself have never been described. We detected the mRNA encoding multiple-splice variants of the GABA(B)R1 and GABA(B)R2 in total RNA isolated from native human and guinea pig airway smooth muscle and from RNA isolated from cultured human airway smooth muscle (HASM) cells. Immunoblots identified the GABA(B)R1 and GABA(B)R2 proteins in human native and cultured airway smooth muscle. The GABA(B)R1 protein was immunohistochemically localized to airway smooth muscle in guinea pig tracheal rings. Baclofen, a GABA(B)R agonist, elicited a concentration-dependent stimulation of [(35)S]GTPgammaS binding in HASM homogenates that was abrogated by the GABA(B)R antagonist CGP-35348. Baclofen also inhibited adenylyl cyclase activity and induced ERK phosphorylation in HASM. Another GABA(B)R agonist, SKF-97541, mimicked while pertussis toxin blocked baclofen's effect on ERK phosphorylation, implicating G(i) protein coupling. Functional GABA(B) receptors are expressed in HASM. GABA may modulate an uncharacterized signaling cascade via GABA(B) receptors coupled to the G(i) protein in airway smooth muscle.


'Lines in the sand': an Australian qualitative study of patient group practices to promote independence from pharmaceutical industry funders.
To study how patient groups that accept pharmaceutical industry money perceive and manage the risk of undue influence from their sponsors. Empirical ethics approach using a qualitative interview study. The Australian patient group sector. 27 participants from 23 patient groups, purposively recruited for diversity of group characteristics (degree of pharmaceutical industry funding, health focus, location) and participant role (staff, board members). Interview data were transcribed and read repeatedly to identify concepts and patterns in the data. These were grouped into conceptual categories that described and explained the findings. We used an inductive analytical approach to identify important themes and concepts in the data. Participants in this study described how the patient group sector receives pressure from pharmaceutical company funders to act in ways that prioritise company interests. Groups worked to try and protect their credibility and ability to act in ways of their own choosing using practical rules or 'lines in the sand' about industry funding activities. They were grouped around the dominant topics of: sponsor exclusivity, brand marketing, agenda setting, advocacy and content of group activities. Lines in the sand were largely experience-driven and ethically informed; they varied between groups. There was also variable transparency among groups about financial interactions with pharmaceutical companies. It is important to know about patient group practices around pharmaceutical industry funders as this allows public scrutiny about the adequacy of such practices. Inadequate strategies may mean that funders can influence patient groups activities in ways that do not necessarily prioritise the interests of members. We found that groups differed in their approach, with little independent external guidance to inform responses to commonly encountered types of influence. Inadequate transparency limits the ability of the public to make informed assessments about the risk of bias over the activities of groups that accept industry funding.


Split and flow: reconfigurable capillary connection for digital microfluidic devices.
Supplying liquid to droplet-based microfluidic microsystems remains a delicate task facing the problems of coupling continuous to digital or macro- to microfluidic systems. Here, we take advantage of superhydrophobic microgrids to address this problem. Insertion of a capillary tube inside a microgrid aperture leads to a simple and reconfigurable droplet generation setup.


Predicting epidemic evolution on contact networks from partial observations.
The massive employment of computational models in network epidemiology calls for the development of improved inference methods for epidemic forecast. For simple compartment models, such as the Susceptible-Infected-Recovered model, Belief Propagation was proved to be a reliable and efficient method to identify the origin of an observed epidemics. Here we show that the same method can be applied to predict the future evolution of an epidemic outbreak from partial observations at the early stage of the dynamics. The results obtained using Belief Propagation are compared with Monte Carlo direct sampling in the case of SIR model on random (regular and power-law) graphs for different observation methods and on an example of real-world contact network. Belief Propagation gives in general a better prediction that direct sampling, although the quality of the prediction depends on the quantity under study (e.g. marginals of individual states, epidemic size, extinction-time distribution) and on the actual number of observed nodes that are infected before the observation time.


[Hygienic aspects of the working conditions assessment and state of health of the workers of the railway departmental security forces].
There were performed a comprehensive hygienic evaluation of the working conditions of employment and analysis of morbidity with temporary disability (MTD) of the workers of the railway departmental security forces. There was proved a significant influence of accumulated period of work on MTD. The professional causality as of the respiratory diseases in shooters and dog handlers, as of diseases of the circulatory system in the administrative and managerial workers has been established.


Investigation of factors controlling PM2.5 variability across the South Korean Peninsula during KORUS-AQ.
The Korea - United States Air Quality Study (May - June 2016) deployed instrumented aircraft and ground-based measurements to elucidate causes of poor air quality related to high ozone and aerosol concentrations in South Korea. This work synthesizes data pertaining to aerosols (specifically, particulate matter with aerodynamic diameters <2.5 micrometers, PM2.5) and conditions leading to violations of South Korean air quality standards (24-hr mean PM2.5 < 35 μg m-3). PM2.5 variability from AirKorea monitors across South Korea is evaluated. Detailed data from the Seoul vicinity are used to interpret factors that contribute to elevated PM2.5. The interplay between meteorology and surface aerosols, contrasting synoptic-scale behavior vs. local influences, is presented. Transboundary transport from upwind sources, vertical mixing and containment of aerosols, and local production of secondary aerosols are discussed. Two meteorological periods are probed for drivers of elevated PM2.5. Clear, dry conditions, with limited transport (Stagnant period), promoted photochemical production of secondary organic aerosol from locally emitted precursors. Cloudy humid conditions fostered rapid heterogeneous secondary inorganic aerosol production from local and transported emissions (Transport/Haze period), likely driven by a positive feedback mechanism where water uptake by aerosols increased gas-to-particle partitioning that increased water uptake. Further, clouds reduced solar insolation, suppressing mixing, exacerbating PM2.5 accumulation in a shallow boundary layer. The combination of factors contributing to enhanced PM2.5 is challenging to model, complicating quantification of contributions to PM2.5 from local versus upwind precursors and production. We recommend co-locating additional continuous measurements at a few AirKorea sites across South Korea to help resolve this and other outstanding questions: carbon monoxide/carbon dioxide (transboundary transport tracer), boundary layer height (surface PM2.5 mixing depth), and aerosol composition with aerosol liquid water (meteorologically-dependent secondary production). These data would aid future research to refine emissions targets to further improve South Korean PM2.5 air quality.


Using oligonucleotide aptamer probes for immunostaining of formalin-fixed and paraffin-embedded tissues.
For tissue immunostaining, antibodies are currently the only clinically validated and commercially available probes. Aptamers, which belong to a class of small molecule ligands composed of short single-stranded oligonucleotides, have emerged as probes over the last several decades; however, their potential clinical value has not yet been fully explored. Using cultured cells and an RNA-based CD30 aptamer, we recently demonstrated that the synthetic aptamer is useful as a specific probe for flow cytometric detection of CD30-expressing lymphoma cells. In this study, we further validated the use of this aptamer probe for immunostaining of formalin-fixed and paraffin-embedded lymphoma tissues. Using CD30 antibody as a standard control, we demonstrated that the synthetic CD30 aptamer specifically recognized and immunostained tumor cells of classical Hodgkin lymphoma and anaplastic large cell lymphoma, but did not react with background cells within tumor sites. Notably, the CD30 aptamer probe optimally immunostained lymphoma cells with lower temperature antigen retrieval (37 vs 96°C for antibody) and shorter probing reaction times (20 vs 90 min for antibody) than typical antibody immunostaining protocols. In addition, the CD30 aptamer probe showed no nonspecific background staining of cell debris in necrotic tissue and exhibited no cross-reaction to tissues that do not express CD30, as confirmed by a standard CD30 antibody staining. Therefore, our findings indicate that the synthetic oligonucleotide CD30 aptamer can be used as a probe for immunostaining of fixed tissue sections for disease diagnosis.


[Experience with one-lung anesthesia using balloon catheter for patients with terminal tracheostomy].
We experienced two cases of lung cancer with terminal tracheostomy. Conventional double lumen tubes such as the Broncho-Cath or the Robertshaw were not applicable for one-lung anesthesia for these cases because of anatomical reasons due to the total laryngectomy. One left lower sleeve lobectomy and a right middle lobectomy were performed under one-lung ventilation using an occlusion balloon catheter that was introduced through a seath placed in the spiral type tracheostomy tube through a plastic connector. This method has three advantages: 1) a spiral type tracheostomy tube is easily fitted for the deformed trachea. 2) an occlusion balloon catheter with an extra lumen at its tip enables the airway aspiration and the inflation of the lung. 3) air leakage around a catheter is completely prevented by the use of a catheter introducer. The present method was concluded to be superior to the prior technique with the Forgaty catheter as an endbronchial blocker.


Associations of snoring frequency with blood pressure among the lean Japanese population: the Toon Health Study.
To date, there are few studies in Asian populations on the association between snoring (a major clinical symptom of sleep apnea) and hypertension. This study aims to examine whether snoring frequency is associated with blood pressure and hypertension in the general Japanese population, after adjustment for major confounding factors. A cross-sectional study of 2021 middle-aged Japanese men and women enrolled in the Toon Health Study between 2009-2012 was conducted. Snoring frequency was assessed using a self-reported questionnaire, and was classified into four categories: never, ≤2 times/week, ≥3 times/week, and unknown. Multivariable regression coefficients for each snoring category were calculated for systolic and diastolic blood pressure, and their odds ratios (ORs) and 95% confidence intervals (95% CIs) for hypertension were calculated after adjusting for major confounding factors. The same analyses were also conducted after stratification by several major confounding factors. Multivariable-adjusted means of systolic and diastolic blood pressure among individuals who snored ≥3 times/week were 4.57 mmHg and 2.58 mmHg higher, respectively, than in individuals who never snored (p < 0.05). The multivariable-adjusted OR (95% CI) for hypertension in the group that snored ≥3 times/week was 1.79 (1.29-2.48), compared with the group that never snored. We also found a significant positive association between snoring frequency and hypertension not only in normal and overweight individuals, but also in lean individuals (body mass index ≤22.8 kg/m2). Higher snoring frequency was associated with higher blood pressure and hypertension among both lean and non-lean Japanese.


Homies with aspirations and positive peer network ties: associations with reduced frequent substance use among gang-affiliated Latino youth.
In marginalized urban neighborhoods across the USA, Latino youth are disproportionately represented among the growing number of youth gangs. Substance use among gang-involved youth poses both immediate and long-term health risks and can threaten educational engagement, future socioeconomic stability, and desistance. Conventional assessments of gang-affiliated youth and their peer network overlook the possibility that positive peer ties may exist and can foster health promoting behavior norms. Drawing on a positive deviance framework, in this study, we examine the relationship between positive peer network characteristics tied to post-secondary educational aspirations and frequent alcohol and marijuana use among Latino, gang-affiliated youth from a neighborhood in San Francisco. Using generalized estimating equations regression models across 72 peer network clusters (162 youth), we found that having close friends who plan to go to a 4-year college was associated with a lower odds of frequent marijuana and alcohol use (OR 0.27, p = 0.02; OR 0.29, p = 0.14, respectively) and that this association persisted when adjusting for risk characteristics (OR 0.19, p < 0.01; OR 0.25, p = 0.12). Public health can advance gang intervention efforts by identifying protective and risk factors associated with non-criminal health outcomes to inform participatory research approaches and asset-based interventions that contribute to building healthy communities.


Recurrence of leprosy cases in the State of Mato Grosso, Central-West Brazil.
To compare new registrations of recurrences of leprosy cases in primary healthcare units (PHUs) and in specialized units in the State of Mato Grosso. This was a cross-sectional study based on all new registrations (N = 323) of recurrences of leprosy cases within the Notifiable Disease Information System (SINAN) between 2004 and 2006 that were made in the State of Mato Grosso, Central-West Brazil. The cases diagnosed were compared regarding sex, age, clinical-laboratory characteristics and geographical distribution among the municipalities. To compare and calculate the proportions of the variables, the chi-square test at the significance level of 5% was used. Among the new registrations of recurrences, 20% were confirmed at specialized units and 80% at PHUs. However, most of the diagnoses at PHUs presented negative bacilloscopy findings (c2 = 12.34; p = 0.002). Seventy-one per cent of the cases were among males; the mean age was 43 years. No differences in the percentages of registrations were observed between the healthcare units with regard to clinical form, operational classification or degree of physical incapacity. Out of the total number of municipalities in the state, 64.7% presented recurrences and these accounted for 6 to 20% of all registrations. The new registrations of cases of recurrence in Mato Grosso were influenced by the diagnoses made at PHUs, thus suggesting that there is a deficiency within the healthcare services in recognizing cases of recurrence.


Expression pattern of HIF1α mRNA in brain, heart and liver tissues of Tibet chicken embryos in hypoxia revealed with quantitative real-time PCR.
The problem of hypoxia adaptation in high altitudes is an unsolved brainteaser in the field of the life sciences. As one of the best chicken breeds with adaptability to highland environment, the Tibet chicken, is genetically different from lowland chicken breeds. It is well known that hypoxia has significant impact on growth by inducing the expression of the alpha subunit of the heterodimeric transcription factor, hypoxia-inducible factor 1 (HIF1α). In the present study, HIF1α expression in brain, heart and liver tissues of the Tibet and Dwarf Recessive White chicken embryos were investigated at the different development stages of days 10, 17 and 20 by real-time PCR, and the expression pattern of HIF1α in chicken embryos of the two chicken breeds incubated under conditions of hypoxia (13% O2) and normoxia (21% O2) was studied. The incubation mortality of the Tibet chicken was lower than the Dwarf Recessive White chicken during the whole incubation in hypoxia, and the mRNA expression of HIF1α had presented the differences in three tissues. The results implied that the hypoxia adaptability of the Tibet chicken embryo was higher than the Dwarf Recessive White chicken, especially in the early stage (day 10) and at a later time (day 20) during the incubation period, but the mechanism of the oxygen transfer of embryos of mountain species is not completely understood, and hypoxia adaptability of the Tibetan chicken remains to be studied further.


Bioenergy Sorghum Crop Model Predicts VPD-Limited Transpiration Traits Enhance Biomass Yield in Water-Limited Environments.
Bioenergy sorghum is targeted for production in water-limited annual cropland therefore traits that improve plant water capture, water use efficiency, and resilience to water deficit are necessary to maximize productivity. A crop modeling framework, APSIM, was adapted to predict the growth and biomass yield of energy sorghum and to identify potentially useful traits for crop improvement. APSIM simulations of energy sorghum development and biomass accumulation replicated results from field experiments across multiple years, patterns of rainfall, and irrigation schemes. Modeling showed that energy sorghum's long duration of vegetative growth increased water capture and biomass yield by ~30% compared to short season crops in a water-limited production region. Additionally, APSIM was extended to enable modeling of VPD-limited transpiration traits that reduce crop water use under high vapor pressure deficits (VPDs). The response of transpiration rate to increasing VPD was modeled as a linear response until a VPD threshold was reached, at which the slope of the response decreases, representing a range of responses to VPD observed in sorghum germplasm. Simulation results indicated that the VPD-limited transpiration trait is most beneficial in hot and dry regions of production where crops are exposed to extended periods without rainfall during the season or to a terminal drought. In these environments, slower but more efficient transpiration increases biomass yield and prevents or delays the exhaustion of soil water and onset of leaf senescence. The VPD-limited transpiration responses observed in sorghum germplasm increased biomass accumulation by 20% in years with lower summer rainfall, and the ability to drastically reduce transpiration under high VPD conditions could increase biomass by 6% on average across all years. This work indicates that the productivity and resilience of bioenergy sorghum grown in water-limited environments could be further enhanced by development of genotypes with optimized VPD-limited transpiration traits and deployment of these crops in water limited growing environments. The energy sorghum model and VPD-limited transpiration trait implementation are made available to simulate performance in other target environments.


Ascorbate-induced oxidative inactivation of Zn2+-glycerophosphocholine cholinephosphodiesterase.
Zn2+-glycerophosphocholine cholinephosphodiesterase, responsible for the conversion of glycerophosphocholine into glycerol and phosphocholine, was inactivated during incubation with ascorbic acid at 38 degrees C. The inclusion of copper ions or Fe2+ accelerated the ascorbate-induced inactivation, with Cu2+ or Cu+ being much more effective than Fe2+, suggestive of ascorbate-mediated oxidation. Dehydroascorbic acid had no effect on the phosphodiesterase, but H2O2 inactivated the enzyme in a concentration-dependent manner. Also, the enzyme was inactivated partially by a superoxide anion-generating system but not an HOCl generator. In support of involvement of H2O2 in the ascorbate action, catalase and superoxide dismutase expressed a complete and a partial protection, respectively. However, hydroxy radical scavengers such as mannitol, benzoate, or dimethyl sulfoxide were incapable of preventing the ascorbate action, excluding the participation of extraneous .OH. Although p-nitrophenylphosphocholine exhibited a modest protection against the ascorbate action, a remarkable protection was expressed by amino acids, especially by histidine. In addition, imidazole, an electron donor, showed a partial protection. Separately, when Cu2+-induced inactivation of the phosphodiesterase was compared with the ascorbate-mediated one, the protection and pH studies indicate that the mechanism for the ascorbate action is different from that for the Cu2+ action. Here, it is proposed that Zn2+-glycerophosphocholine cholinephosphodiesterase is one of brain membrane proteins susceptible to oxidative inactivation.


Stimulating natural killer cells to protect against cancer: recent developments.
Current cancer immunotherapies have begun to target cell types involved in innate immunity, such as natural killer (NK) cells that recognize and kill tumor cells. Recent advances in the study of NK cell biology have generated interest in manipulating these cells to generate anti-tumor responses. A rise in the number of activated NK cells has been shown to prevent and treat cancer in many preclinical models and is a positive clinical factor in human tumors. This article will focus on recent research on the ability of IL-15 and Toll-like receptor ligands to stimulate NK cell activity against cancer. The potential of these therapies, both alone and in conjunction with traditional and other vaccine platforms, will be reviewed. The current status of these therapies in clinical trials will also be discussed. Targeting these cell types in the context of human cancers may be an essential factor in future cancer treatments.


Human neutrophils secrete bioactive paucimannosidic proteins from azurophilic granules into pathogen-infected sputum.
Unlike plants and invertebrates, mammals reportedly lack proteins displaying asparagine (N)-linked paucimannosylation (mannose(1-3)fucose(0-1)N-acetylglucosamine(2)Asn). Enabled by technology advancements in system-wide biomolecular characterization, we document that protein paucimannosylation is a significant host-derived molecular signature of neutrophil-rich sputum from pathogen-infected human lungs and is negligible in pathogen-free sputum. Five types of paucimannosidic N-glycans were carried by compartment-specific and inflammation-associated proteins of the azurophilic granules of human neutrophils including myeloperoxidase (MPO), azurocidin, and neutrophil elastase. The timely expressed human azurophilic granule-resident β-hexosaminidase A displayed the capacity to generate paucimannosidic N-glycans by trimming hybrid/complex type N-glycan intermediates with relative broad substrate specificity. Paucimannosidic N-glycoepitopes showed significant co-localization with β-hexosaminidase A and the azurophilic marker MPO in human neutrophils using immunocytochemistry. Furthermore, promyelocyte stage-specific expression of genes coding for paucimannosidic proteins and biosynthetic enzymes indicated a novel spatio-temporal biosynthetic route in early neutrophil maturation. The absence of bacterial exoglycosidase activities and paucimannosidic N-glycans excluded exogenous origins of paucimannosylation. Paucimannosidic proteins from isolated and sputum neutrophils were preferentially secreted upon inoculation with virulent Pseudomonas aeruginosa. Finally, paucimannosidic proteins displayed affinities to mannose-binding lectin, suggesting immune-related functions of paucimannosylation in activated human neutrophils. In conclusion, we are the first to document that human neutrophils produce, store and, upon activation, selectively secrete bioactive paucimannosidic proteins into sputum of lungs undergoing pathogen-based inflammation.


ECAP spread of excitation with virtual channels and physical electrodes.
The primary goal of this study was to evaluate physiological spatial excitation patterns for stimulation of adjacent physical electrodes and intermediate virtual channels. Two experiments were conducted that utilized electrically evoked compound action potential (ECAP) spread-of-excitation (SOE) functions obtained with the traditional forward-masking subtraction method. These two experiments examined spatial excitation patterns for virtual-channel maskers and probes, respectively. In Experiment 1, ECAP SOE patterns were obtained for maskers applied to physical electrodes and virtual channels to determine whether virtual-channel maskers yield SOE patterns similar to those predicted from physical electrodes. In Experiment 2, spatial separation of SOE functions was compared for two adjacent physical probe electrodes and the intermediate virtual channel to determine the extent to which ECAP SOE patterns for virtual-channel probes are spatially separate from those obtained with physical electrodes. Data were obtained for three electrode regions (basal, middle, apical) for 35 ears implanted with Cochlear (N = 16) or Advanced Bionics (N = 19) devices. Results from Experiment 1 showed no significant difference between predicted and measured ECAP amplitudes for Advanced Bionics subjects. Measured ECAP amplitudes for virtual-channel maskers were significantly larger than the predicted amplitudes for Cochlear subjects; however, the difference was <2 μV and thus is likely not clinically significant. Results from Experiment 2 showed that the probe set in the apical region demonstrated the least amount of spatial separation amongst SOE functions, which may be attributed to more uniform nerve survival patterns, closer electrode spacing, and/or the tapered geometry of the cochlea. As expected, adjacent physical probes demonstrated greater spatial separation than for comparisons between each physical probe and the intermediate virtual channel. Finally, the virtual-channel SOE functions were generally weighted toward the basal electrode in the pair.


Exploring the effectiveness of novel benzimidazoles as CB2 ligands: synthesis, biological evaluation, molecular docking studies and ADMET prediction.
Herein we continued our previous work on the development of CB2 ligands, reporting the design and synthesis of a series of benzimidazole-containing derivatives that were explored as selective CB2 ligands with binding affinity towards both CB1 and CB2 receptors. Seven out of eighteen compounds exhibited preferential binding ability to CB2 over CB1 receptors with potencies in the sub-micromolar or low micromolar range. In particular, we identified two promising hit compounds, the agonist 1-[2-(N,N-diethylamino)ethyl]-2-(4-ethoxybenzyl)-5-trifluoromethylbenzimidazole (3) (CB2: K i = 0.42 μM) and the inverse agonist/antagonist 1-butyl-2-(3,4-dichlorobenzyl)-5-trifluoromethylbenzimidazole (11) (CB2: K i = 0.37 μM). Docking studies also performed on other benzimidazoles reported in the literature supported the structure-activity relationship observed in this series of compounds and allowed the key contacts involved in the agonist and/or inverse agonist behaviour displayed by these derivatives to be determined. The in silico evaluation of ADMET properties suggested a favorable pharmacokinetic and safety profile, promoting the drug-likeness of these compounds towards a further optimization process.


Denitrovibrio acetiphilus, a novel genus and species of dissimilatory nitrate-reducing bacterium isolated from an oil reservoir model column.
A novel dissimilatory, nitrate-reducing bacterium, designated strain N2460T, was isolated from an oil reservoir model column. Strain N2460T is a mesophilic, obligately anaerobic, marine, gram-negative bacterium. The cells are vibrio-shaped and motile by a bipolar flagellum. Strain N2460T reduces nitrate to ammonia in a mineral medium supplied by acetate. The presence of a 2-oxoglutarate dehydrogenase activity indicates that acetate is oxidized via the citric acid cycle. No growth is obtained on formate, higher fatty acids, malate, fumarate, benzoate, alcohols, sugar, yeast extract, crude oil, alkanes, proline, hydrogen, sulfur or thiosulfate with nitrate as electron acceptor. Oxygen, sulfate, thiosulfate and sulfur are not utilized as alternative electron acceptors. Strain N2460T grows fermentatively on fumarate, but not on pyruvate. The G+C content of the DNA is 42.6 mol%. 16S rRNA gene analysis shows that strain N2460T belongs to the Bacteria and that the closest relative is 'Geovibrio ferrireducens' (sequence similarity 86.9%). On the basis of phylogenetic as well as phenotypic data, it is proposed that strain N2460T represents the type strain of a new genus and species, Denitrovibrio acetiphilus gen. nov., sp. nov.


Defining competition in markets: why and how?
To examine the variety of perspectives from which to study the measurement of competition in the healthcare marketplace. Based on a meeting held by The Robert Wood Johnson Foundation in 1996, the authors discuss the complications inherent in the way markets and products are defined by key stakeholders, including economists, policymakers, federal antitrust officials, purchasers, and the competitors themselves. The consensus among those who study this issue is that the way competitors, markets, and geographic areas are currently defined, and the ways of measuring competition, are inadequate, due mainly to the fact that both the measures and the definitions have been constructed from very limited data. Confounding this is the fact that analyses of competition are undertaken for such a wide variety of uses and that creating one database to solve the problems mentioned can be extremely daunting. Future research should examine ways to develop better definitions of the new healthcare structures that are competing with each other and ways to create measures of competition that include these new structures. To remedy gaps in the ability to measure competition, the field might also benefit from a public use data file, similar to the Area Resource File (ARF), that would contain HMO data according to geographic area, as well as provider data, employer data, payer data, and sociodemographic data.


Atmospheric pollution with halothane during outpatient dental anaesthesia.
Pollution of the atmosphere with halothane has been measured during general anaesthesia for outpatients undergoing dental extractions. The level of contamination was far in excess of that recorded in surgical operating theatres, and halothane was inhaled at similar concentrations by both the anaesthetist and the dental surgeon. The room air-changing system and a local fan had little effect on contamination. Scavenging devices on the exhalation valve of the anaesthetic circuit were only moderately effective in reducing the concentration of halothane in the atmosphere.


Sensitive and selective determination of tetracaine and its metabolite in human samples by gas chromatography-mass spectrometry.
A sensitive and reliable method was developed for the determination of tetracaine and its metabolite, p-butylaminobenzoic acid, in human samples. Tetracaine and the metabolite, effectively extracted using a liquid-liquid extraction procedure from 0.5 g of sample, were analyzed by gas chromatography-mass spectrometry. Tetracaine was analyzed without derivatization, and the metabolite was analyzed after tert-butylolimethylsilyl derivatization. Dibucaine and p-dimethylaminobenzoic acid were used as internal standards for tetracaine and the metabolite, respectively. The calibration curve for each compound was linear in the concentration range from 10 to 1,000 ng/0.5 g, and the lower limits of detection were 10 ng/g for tetracaine and 0.6 ng/g for the metabolite in whole blood and tissues. The accuracy and precision of the method were evaluated in whole blood and brain at the concentrations of 50 ng/0.5 g and 500 ng/0.5 g for tetracaine and 10 ng/0.5 g and 100 ng/0.5 g for the metabolite. The coefficients of variation ranged from 0.8 to 3.0% for tetracaine and 2.4 to 9.8% for the metabolite. We used this method to determine tetracaine and its metabolite in human whole blood and tissues of an autopsied patient who died during spinal anesthesia induced by tetracaine.


Changes in physiological and behavioural pain indicators over time in preterm and term infants at risk for neurologic impairment.
Approximately 10% of infants admitted to a Neonatal Intensive Care Unit (NICU) are at risk for Neurological Impairment (NI). While we have limited knowledge on the influence of NI risk on pain responses, we have no knowledge of how these responses change over time. To compare physiological and behavioural pain responses of infants at three levels of NI risk during the NICU neonatal period (Session 1) and at 6 months of age (Session 2). Prospective observational design with 149 preterm and term infants at high (Cohort A, n=54), moderate (Cohort B, n=45) and mild (Cohort C, n=50) risks for NI from 3 Canadian tertiary level NICUs. Infants were observed in the NICU during 3 standardized phases of a heel lance: baseline, stick and return-to-baseline. At 6 months, infants were observed during the same three phases during an intramuscular immunization injection. Physiological (heart rate, oxygen saturation) and behavioural (9 facial actions, cry) responses were continuously recorded. A significant interaction of Phase by Session was found with less total facial activity observed during Session 2 (all p values<0.04). A significant interaction for Session by Cohort was found, showing that infants in Cohort A had significantly more change from baseline-to-stick phase for brow bulge, eye squeeze, nasolabial furrow and open lips between sessions with less facial actions demonstrated at Session 2 (all p<0.02). There were significantly lower mean and minimum heart rate (all p<0.02) and higher minimum and maximum oxygen saturation (p<0.04) at Session 2. Significantly higher mean and minimum fundamental cry frequencies (pitch) in Cohort B (p<0.04) were found in Session 1. Cohort A had significantly longer cry durations, but no significant differences in cry dysphonation. Behavioural and physiological infant pain responses were generally diminished at 6 months of age compared to those in the neonatal period with some differences between NI risk groups in cry responses. Future exploration into the explanation for these differences between sessions and cohorts is warranted.


Germplasm Resources and Metabolite Marker Screening of High-Flavonoid Tartary Buckwheat (Fagopyrum tataricum).
Tartary buckwheat is an annual minor cereal crop with a variety of secondary metabolites, endowing it with a high nutritional and medicinal value. Flavonoids constitute the primary compounds of Tartary buckwheat. Recently, metabolomics, as an adjunct breeding method, has been increasingly employed in crop research. This study explores the correlation between the total flavonoid content (TFC) and antioxidant capacity in 167 Tartary buckwheat varieties. Ten Tartary buckwheat varieties with significant differences in flavonoid content and antioxidant capacity were selected by cluster analysis. With the use of liquid chromatography-mass spectrometry, 58 flavonoid compounds were identified, namely, 42 flavonols, 10 flavanols, 3 flavanones, 1 isoflavone, 1 anthocyanidin, and 1 proanthocyanidin. Different samples were clearly separated by employing principal component analysis and partial least-squares discriminant analysis. Eight differential flavonoid compounds were further selected through volcano plots and variable importance in projection. Differential metabolites were highly correlated with TFC and antioxidant capacity. Finally, metabolic markers of kaempferol-3-O-hexoside, kaempferol-7-O-glucoside, and naringenin-O-hexoside were determined by the random forest model. The findings provide a basis for the selection and identification of Tartary buckwheat varieties with high flavonoid content and strong antioxidant activity.


Genome-wide significant localization for working and spatial memory: Identifying genes for psychosis using models of cognition.
It is well established that risk for developing psychosis is largely mediated by the influence of genes, but identifying precisely which genes underlie that risk has been problematic. Focusing on endophenotypes, rather than illness risk, is one solution to this problem. Impaired cognition is a well-established endophenotype of psychosis. Here we aimed to characterize the genetic architecture of cognition using phenotypically detailed models as opposed to relying on general IQ or individual neuropsychological measures. In so doing we hoped to identify genes that mediate cognitive ability, which might also contribute to psychosis risk. Hierarchical factor models of genetically clustered cognitive traits were subjected to linkage analysis followed by QTL region-specific association analyses in a sample of 1,269 Mexican American individuals from extended pedigrees. We identified four genome wide significant QTLs, two for working and two for spatial memory, and a number of plausible and interesting candidate genes. The creation of detailed models of cognition seemingly enhanced the power to detect genetic effects on cognition and provided a number of possible candidate genes for psychosis.


Deconvolution based photoacoustic reconstruction with sparsity regularization.
In most photoacoustic tomography (PAT) reconstruction approaches, it is assumed that the receiving transducers have omnidirectional response and can fully surround the region of interest. These assumptions are not satisfied in practice. To deal with these limitations, we present a novel deconvolution based photoacoustic reconstruction with sparsity regularization (DPARS) technique. The DPARS algorithm is a semi-analytical reconstruction approach in which the projections of the absorber distribution derived from a deconvolution-based method are computed and used to generate a large linear system of equations. In these projections, computed over limited viewing angles, the directivity effect of the transducer is taken into account. The distribution of absorbers is computed using a sparse representation of absorber coefficients obtained from the discrete cosine transform. This sparse representation helps improve the numerical conditioning of the system of equations and reduces the computation time of the deconvolution-based approach by one order of magnitude relative to Tikhonov regularization. The algorithm has been tested in simulations, and using two-dimensional and three-dimensional experimental data obtained with a conventional ultrasound transducer. The results show that DPARS, when evaluated using contrast-to-noise ratio and root-mean-square errors, outperforms the conventional delay-and-sum (DAS) reconstruction method.


Multimodality Image-Guided Cryoablation for Inoperable Tumor-Induced Osteomalacia.
Tumor-induced osteomalacia (TIO) is a debilitating paraneoplastic condition caused by small phosphaturic mesenchymal tumors (PMTs) that secrete large amounts of the phosphate-regulating and vitamin D-regulating hormone, FGF23. Tumor removal results in cure. However, because of high perioperative comorbidity, either from tumor location or host factors, surgery is sometimes not an option. Tumor destruction via cryoablation may be an effective option for inoperable PMTs. Three subjects with a confirmed diagnosis of TIO were studied. All three underwent cryoablation of suspected PMTs rather than surgery due to significant medical comorbidities or challenging anatomical location. Subject 3 had tumor embolization 24 hours prior to cryoablation because of the size and hypervascularity of the tumor. The success of the tumor cryoablation was defined by normalization of serum phosphate and FGF23. Cryoablation resulted in a rapid decrease in plasma intact FGF23 by 24 hours postprocedure in all three subjects (0, 2, and 9 pg/mL, respectively) with normalization of blood phosphate by postprocedure day 3. Three-day renal tubular reabsorption of phosphate increased to 76%, 94%, and 95.2%, respectively; 1, 25(OH)2 vitamin D increased to 84, 138, and 196 pg/ml, respectively. All three had dramatic clinical improvement in pain and weakness. Two subjects tolerated the procedure well with no complications; one had significant prolonged procedure-related localized pain. Although surgery remains the treatment of choice, cryoablation may be an effective, less invasive, and safe treatment for patients with difficult to remove tumors or who are poor surgical candidates. © 2017 American Society for Bone and Mineral Research.


Bird expertise does not increase motion sensitivity to bird flight motion.
While motion information is important for the early stages of vision, it also contributes to later stages of object recognition. For example, human observers can detect the presence of a human, judge its actions, and judge its gender and identity simply based on motion cues conveyed in a point-light display. Here we examined whether object expertise enhances the observer's sensitivity to its characteristic movement. Bird experts and novices were shown point-light displays of upright and inverted birds in flight, or upright and inverted human walkers, and asked to discriminate them from spatially scrambled point-light displays of the same stimuli. While the spatially scrambled stimuli retained the local motion of each dot of the moving objects, it disrupted the global percept of the object in motion. To estimate a detection threshold in each object domain, we systematically varied the number of noise dots in which the stimuli were embedded using an adaptive staircase approach. Contrary to our predictions, the experts did not show disproportionately higher sensitivity to bird motion, and both groups showed no inversion cost. However, consistent with previous work showing a robust inversion effect for human motion, both groups were more sensitive to upright human walkers than their inverted counterparts. Thus, the result suggests that real-world experience in the bird domain has little to no influence on the sensitivity to bird motion and that birds do not show the typical inversion effect seen with humans and other terrestrial movement.


The Extracellular IFI16 Protein Propagates Inflammation in Endothelial Cells Via p38 MAPK and NF-κB p65 Activation.
The nuclear interferon-inducible-16 (IFI16) protein acts as DNA sensor in inflammasome signaling and as viral restriction factor. Following Herpesvirus infection or UV-B treatment, IFI16 delocalizes from the nucleus to the cytoplasm and is eventually released into the extracellular milieu. Recently, our group has demonstrated the occurrence of IFI16 in sera of systemic-autoimmune patients that hampers biological activity of endothelia through high-affinity membrane binding. As a continuation, we studied the activity of endotoxin-free recombinant IFI16 (rIFI16) protein on primary endothelial cells. rIFI16 caused dose/time-dependent upregulation of IL-6, IL-8, CCL2, CCL5, CCL20, ICAM1, VCAM1, and TLR4, while secretion of IL-6 and IL-8 was amplified with lipopolysaccharide synergy. Overall, cytokine secretion was completely inhibited in MyD88-silenced cells and partially by TLR4-neutralizing antibodies. By screening downstream signaling pathways, we found that IFI16 activates p38, p44/42 MAP kinases, and NF-kB. In particular, activation of p38 is an early event required for subsequent p44/42 MAP kinases activity and cytokine induction indicating a key role of this kinase in IFI16 signaling. Altogether, our data conclude that extracellular IFI16 protein alone or by synergy with lipopolysaccharide acts like Damage-associated molecular patterns propagating "Danger Signal" through MyD88-dependent TLR-pathway.


Understanding discharge communication behaviours in a pediatric emergency care context: a mixed methods observation study protocol.
One of the most important transitions in the continuum of care for children is discharge to home. Optimal discharge communication between healthcare providers and caregivers (e.g., parents or other guardians) who present to the emergency department (ED) with their children is not well understood. The lack of policies and considerable variation in practice regarding discharge communication in pediatric EDs pose a quality and safety risk for children and their parents. The aim of this mixed methods study is to better understand the process and structure of discharge communication in a pediatric ED context to contribute to the design and development of discharge communication interventions. We will use surveys, administrative data and real-time video observation to characterize discharge communication for six common illness presentations in a pediatric ED: (1) asthma, (2) bronchiolitis, (3) abdominal pain, (4) fever, (5) diarrhea and vomiting, and (6) minor head injury. Participants will be recruited from one of two urban pediatric EDs in Canada. Video recordings will be analyzed using Observer XT. We will use logistic regression to identify potential demographic and visit characteristic cofounders and multivariate logistic regression to examine association between verbal and non-verbal behaviours and parent recall and comprehension. Video recording of discharge communication will provide an opportunity to capture important data such as temporality, sequence and non-verbal behaviours that might influence the communication process. Given the importance of better characterizing discharge communication to identify potential barriers and enablers, we anticipate that the findings from this study will contribute to the development of more effective discharge communication policies and interventions.


Quercetin inhibits inflammatory bone resorption in a mouse periodontitis model.
Periodontitis is a disease that leads to bone destruction and represents the main cause of tooth loss in adults. The development of aggressive periodontitis has been associated with increased inflammatory response that is induced by the presence of a subgingival biofilm containing Aggregatibacter actinomycetemcomitans. The flavonoid quercetin (1) is widespread in vegetables and fruits and exhibits many biological properties for possible medical and clinical applications such as its anti-inflamatory and antioxidant effects. Thus, in the present study, the properties of 1 have been evaluated in bone loss and inflammation using a mouse periodontitis model induced by A. actinomycetemcomitans infection. Subcutaneous treatment with 1 reduced A. actinomycetemcomitans-induced bone loss and IL-1β, TNF-α, IL-17, RANKL, and ICAM-1 production in the gingival tissue without affecting bacterial counts. These results demonstrated that quercetin exhibits protective effects in A. actinomycetemcomitans-induced periodontitis in mice by modulating cytokine and ICAM-1 production.


A Systematic Review of 10 Years of Augmented Reality Usability Studies: 2005 to 2014.
Augmented Reality (AR) interfaces have been studied extensively over the last few decades, with a growing number of user-based experiments. In this paper, we systematically review 10 years of the most influential AR user studies, from 2005 to 2014. A total of 291 papers with 369 individual user studies have been reviewed and classified based on their application areas. The primary contribution of the review is to present the broad landscape of user-based AR research, and to provide a high-level view of how that landscape has changed. We summarize the high-level contributions from each category of papers, and present examples of the most influential user studies. We also identify areas where there have been few user studies, and opportunities for future research. Among other things, we find that there is a growing trend toward handheld AR user studies, and that most studies are conducted in laboratory settings and do not involve pilot testing. This research will be useful for AR researchers who want to follow best practices in designing their own AR user studies.


One Outcome, Many Trends: Understanding National Data Sources for Road Traffic Fatalities in China.
To better understand national data sources and evaluate time trends in road traffic fatalities (RTF) in China. We reviewed national sources on RTF data. These included population-based report from the Ministry of Public Security (MPS), sample-based estimates from the Ministry of Health (MOH) and the Disease Surveillance Points System (DSP), as well as model-based estimates from the World Health Organization (WHO), and the Global Burden of Disease Study (GBD). All data sources have limitations in coverage, representativeness or overreliance on model specifications. Despite the discrepancies in methodologies and estimates, all sources indicated an increase in RTF before 2005. Since then, MPS and GBD indicated a decrease, DSP showed plateauing, and MOH and WHO suggested increasing fatalities. However, despite any recent decline, RTF remain high. The divergent trends in RTF across data sets in China implies a challenge to understanding China's experience with addressing RTF. China needs to reconcile data sources and further improve road safety.


Alkaloids in marine algae.
This paper presents the alkaloids found in green, brown and red marine algae. Algal chemistry has interested many researchers in order to develop new drugs, as algae include compounds with functional groups which are characteristic from this particular source. Among these compounds, alkaloids present special interest because of their pharmacological activities. Alkaloid chemistry has been widely studied in terrestrial plants, but the number of studies in algae is insignificant. In this review, a detailed account of macro algae alkaloids with their structure and pharmacological activities is presented. The alkaloids found in marine algae may be divided into three groups: 1. Phenylethylamine alkaloids, 2. Indole and halogenated indole alkaloids, 3. Other alkaloids.


Control of Alicyclobacillus acidoterrestris in fruit juices by a newly discovered bacteriocin.
Alicyclobacillus acidoterrestris is one of the most spoilage-causing bacteria in fruit juices. Control of A. acidoterrestris in fruit juices by bificin C6165 (Pei et al. in J Appl Microbiol 114(5):1273-1284, 2013), a bacteriocin produced by Bifidobacterium animalis subsp. animalis CICC 6165, was described in this study. Activity spectrum of bificin C6165 was investigated and sixteen strains of A. acidoterrestris were sensitive to bificin C6165 in diluted Apple Juices. In the commercial fruit juices, vegetative cells of A. acidoterrestris were inactivated by bificin C6165 at 40 μg/ml. The inhibitory effect of bificin C6165 was better at lower pH (pH 3.5) and at a higher temperature of 45 °C. Furthermore, electron microscopy examination of the vegetative cells treated with bacteriocin revealed substantial cell damage and bacterial lysis. The result suggested that primary mode of action of bificin C6165 was most probably due to pore formation. Although no significantly activity of bificin C6165 was observed against the endospores of A. acidoterrestris in commercial apple juice, the addition of bacteriocin contributed to the reduction of the thermal resistance of A. acidoterrestris spores. Additionally, encapsulation of bificin C6165 with Ca-alginate gel was investigated. Encapsulation of bificin C6165 provided a promising method to control A. acidoterrestris in food juice industry.


Osteomalacia after Roux-en-Y gastric bypass.
To emphasize the potential for Roux-en-Y gastric bypass treatment of morbid obesity to result in late development of metabolic bone disease and to illustrate the error of treating a low bone mineral density with bisphosphonates in the presence of unrecognized osteomalacia. We conducted a retrospective case review of clinical, laboratory, and radiologic details in a patient who underwent Roux-en-Y gastric bypass as well as a review of the literature relative to metabolic bone disease associated with bariatric surgical procedures. A 42-year-old woman was diagnosed with high bone turnover osteoporosis and failed to respond to bisphosphonate (alendronate) therapy. Her past medical history included corticosteroid-dependent asthma and a Roux-en-Y gastric bypass surgical procedure for obesity approximately 6 1/2 years before the current assessment. Evaluation revealed vitamin D deficiency in conjunction with pronounced secondary hyperparathyroidism and biochemical evidence of osteomalacia. Aggressive calcium and vitamin D supplementation corrected the vitamin D-deficient state and was accompanied by rapid improvement in clinical symptoms, biochemical variables, and bone mineral density. This case exemplifies two principles: (1) the potential for a Roux-en-Y gastric bypass surgical procedure to lead to the development of metabolic bone disease and (2) the importance of recognizing mineralization defects as a cause for low bone mineral density, before initiation of therapy with bisphosphonates.


Glucocorticoid response to both predictable and unpredictable challenges detected as corticosterone metabolites in collared flycatcher droppings.
In most vertebrate animals, glucocorticoid hormones are the chief mediators of homeostasis in response to ecological conditions and as they progress through their lifecycle. In addition, glucocorticoids are a major part of the stress response and stress induced elevations of the hormone can make it difficult to assess glucocorticoid secretion in response to changes in life-stage and current environmental conditions in wild animals. Particularly when quantifying circulating levels of glucocorticoids in the blood which fluctuate rapidly in response to stress. An alternative method of quantifying glucocorticoids is as hormone metabolites in faeces or urine giving a historical sample related to the gut passage time and urinary tract that is less sensitive to stressful events which cause spikes in the circulating hormone level. Although the concentration of glucocorticoid metabolites are influenced by faecal mass thereby potentially affecting any differences in hormone metabolites detected amongst samples. In the present study, we aimed to detect changes in levels of corticosterone, the primary bird glucocorticoid, in relation to the phase of reproduction, in a breeding population of collared flycatchers by sampling corticosterone metabolites in droppings. We also tested how corticosterone metabolite concentrations were affected by ambient temperature and related to body condition in adult birds. Our results indicate that the upregulation of corticosterone between incubation and nestling feeding in female birds is crucial for successful reproduction in this species. Also, females appear to downregulate corticosterone during incubation in response to lower ambient temperature and poorer body condition. Our results did not indicate a relationship between dropping mass and corticosterone metabolite concentrations, which suggests that our findings were linked to the regulation of corticosterone in response to predictable and unpredictable challenges.


[Value of various concepts in long-term electrocardiography].
With the introduction of long-term ECG by N. Holter more than 25 years ago a more sensitive detection of spontaneous arrhythmias and ST-changes became possible compared with the standard ECG at rest. Unfortunately, visual high-speed analysis of the tapes caused remarkable loss of stored informations due to the failure of rapid identification. Full disclosure of the ECG, therefore, is a valuable supplement to long-term ECG. As an isolated method, however, it cannot be recommended since the significantly reduced amplitude and paper speed of full disclosure likewise is responsible for a remarkable loss of informations. Only computer-aided analysis of the ECG provides more complete information and at the same time more quantitative data. Analysis of the accuracy of different computer-based systems showed, however, that a sensitivity as well as a positive predictive accuracy of 100% remains the exception to the rule. Moreover, the spectrum of cardiac arrhythmias and ST-changes identifiable by automatic analysis is small. For these reasons, interaction of the user is mandatory--in order to validate computer results as well as to detect ECG abnormalities unidentifiable by computer. The first is possible only to a limited degree with discontinuous ECG recording by event recorders, the last remains impossible. Thus, the promising concept of real-time cardiac monitors should be considered investigational or experimental at this time.


Synthesis of delta-tributylstannyl-alpha,beta,gamma,delta-unsaturated aldehydes from pyridines.
Zincke aldehydes, which are readily available from the ring-opening reaction of pyridinium salts, are easily converted into delta-tributylstannyl-alpha,beta,gamma,delta-unsaturated aldehydes (stannyldienals) by the action of tributylstannyllithium. This reaction appears to proceed via 1,6-stannyllithium addition/elimination of lithium dialkylamide. Several stannyldienals of significant utility for the synthesis of polyene natural products have been made by this route, which proceeds in modest yields, but is successful on multigram scale using inexpensive reagents. Simple stannylenals and stannylenones are similarly available from the corresponding vinylogous amides.


Novobiocin inhibits interactions required for yeast TFIIIB sequestration during stable transcription complex formation in vitro.
Novobiocin concentrations normally used to inhibit a putative eukaryotic DNA gyrase have been found to inhibit transcription of a yeast 5S rRNA gene using an in vitro yeast transcription system. Purified RNA polymerase III and three yeast transcription factors (chromatographically separated, partially purified and free of any detectable gyrase activity) were used. Novobiocin prevents specific transcription if added to the in vitro system immediately prior to the addition of transcription factors and RNA polymerase. If a stable transcription factor complex is allowed to form prior to the addition of novobiocin, concentrations of novobiocin as high as 1000 micrograms/ml have no effect on in vitro transcription. Transcription factors TFIIIA and TFIIIC are able to be stably sequestered onto 5SrDNA-cellulose, but factor TFIIIB is not able to associate with the 5SrDNA-TFIIIA-TFIIIC complex in the presence of novobiocin. Although novobiocin is able to precipitate other basic proteins, it does not appear to precipitate any of these class III gene transcription factors, but instead appears to act by disrupting specific factor-factor interactions.


Pot size matters: a meta-analysis of the effects of rooting volume on plant growth.
The majority of experiments in plant biology use plants grown in some kind of container or pot. We conducted a meta-analysis on 65 studies that analysed the effect of pot size on growth and underlying variables. On average, a doubling of the pot size increased biomass production by 43%. Further analysis of pot size effects on the underlying components of growth suggests that reduced growth in smaller pots is caused mainly by a reduction in photosynthesis per unit leaf area, rather than by changes in leaf morphology or biomass allocation. The appropriate pot size will logically depend on the size of the plants growing in them. Based on various lines of evidence we suggest that an appropriate pot size is one in which the plant biomass does not exceed 1gL-1. In current research practice ~65% of the experiments exceed that threshold. We suggest that researchers need to carefully consider the pot size in their experiments, as small pots may change experimental results and defy the purpose of the experiment.


The relationship between time from myocardial infarction, left ventricular dyssynchrony, and the risk for ventricular arrhythmia: speckle-tracking echocardiographic analysis.
Differences in arrhythmogenic substrate may explain the variable efficacy of implantable cardioverter-defibrillators (ICDs) in primary sudden cardiac death prevention over time after myocardial infarction (MI). Speckle-tracking echocardiography allows the assessment left ventricular (LV) dyssynchrony, which may reflect the electromechanical heterogeneity of myocardial tissue. The aim of the present study was to evaluate the relationship among LV dyssynchrony, age of MI, and their association with the risk for ventricular tachycardia (VT) after MI. A total of 206 patients (median age, 67 years; 87% men) with prior MIs (median MI age, 6.2 years; interquartile range, 0.66-15 years) who underwent programmed electrical stimulation, speckle-tracking echocardiography, and ICD implantation were retrospectively evaluated. LV dyssynchrony was defined as the standard deviation of time to peak longitudinal systolic strain values using speckle-tracking strain echocardiography. LV scar burden was evaluated by the percentage of segments exhibiting scar (defined as an absolute longitudinal strain of magnitude < 4.5%). Patients were followed up for the occurrence of first monomorphic VT requiring ICD therapy (antitachycardia pacing or shock) for a median of 24 months. In total, 75 individuals experienced the primary end point of monomorphic VT. LV dyssynchrony was independently associated with the occurrence of VT at follow-up (hazard ratio per 10-msec increase, 1.12; 95% confidence interval, 1.07-1.18; P < .001), together with nonrevascularization of the infarct-related artery and VT inducibility. Patients with older (>180 months) MIs had a higher likelihood of VT inducibility (88% vs 63%, P = .003) and greater scar burden (14.7 ± 15.8% vs 10.7 ± 11.4%, P = .03) compared with patients with recent (<8 months) MIs. LV dyssynchrony is independently associated with the occurrence of VT after MI.


[Expectations of nursing students from their initial hospital training experiences].
The first experience in hospital training may create tensions and anxieties. These feelings interfere negatively in the teaching-learning process. The aim of this paper was to investigate the expectations of Nursing Fundamentals II students regarding their initial hospital training. Second-year students of the Nursing Course offered by Universidade Estadual de Maringá (state of Paraná, Brazil) were chosen as subjects of this investigation. The students revealed a state of anxiety when confronting technical procedures, relationship with patients and evolution. By reflecting on these aspects instructor the may increase the teaching-learning efficiency and humanize the professional training.


Molecular identification by generating coherence between molecular normal modes using stimulated Raman scattering.
We propose a method of driving the vibrations of normal modes of a target molecule into coherence using stimulated Raman scattering. In concert many vibrations can produce a larger anti-Stokes signal than a single vibration. The same illumination does not drive other molecules to have coherent vibrations so that these molecules produce a weaker signal. We investigate how target and confounder molecules can be distinguished by pulses that drive many vibrations coherently, which has applications in coherent Raman microspectroscopy.


Comparative ceftaroline activity tested against pathogens associated with community-acquired pneumonia: results from an international surveillance study.
To document the spectrum of activity of ceftaroline, the active form of the prodrug, ceftaroline fosamil, a new cephalosporin with anti-methicillin-resistant staphylococcal activity, against a surveillance collection of clinical isolates obtained from the USA and Europe during 2008-09. A selected group of species associated with community-acquired pneumonia (CAP; 6496 of 17 326 monitored strains) were tested for susceptibility in a central laboratory using CLSI broth microdilution methods. Organisms were sampled from 55 medical centres, 27 in the USA and 28 (12 countries) in Europe. Ceftaroline and comparator agents were tested and interpretations of MIC endpoints made by applying current CLSI (2010) and EUCAST (2010) breakpoint criteria. Against 1340 Streptococcus pneumoniae, ceftaroline inhibited all isolates at ≤0.5 mg/L (MIC(50/90), ≤0.008/0.12 mg/L) and was 8-fold more active than ceftriaxone (MIC(90), 1 mg/L; only 79.2% coverage at EUCAST breakpoint). Haemophilus influenzae (n = 584; MIC(50/90), ≤0.008/0.015 mg/L), Moraxella catarrhalis (n = 377; MIC(50/90), 0.03-0.06/0.12 mg/L) and Staphylococcus aureus (n = 590; MIC(50/90), 0.5/1 mg/L) were very susceptible to ceftaroline, regardless of β-lactamase production or multidrug resistance (MDR) patterns. The potency of ceftaroline against three species of Enterobacteriaceae (Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae) was similar to that of ceftriaxone, ceftazidime and piperacillin/tazobactam. Only modest differences in rates of ceftaroline susceptibility (breakpoint ≤2 mg/L) were noted with extended-spectrum β-lactamase-negative Enterobacteriaceae strains between the USA and Europe (97.9% versus 97.0% for E. coli). Ceftaroline, like ceftriaxone, was not active against ceftazidime-resistant E. coli (10.2%-26.2% susceptible at ≤2 mg/L) or K. pneumoniae (5.3%-11.2%). The ceftaroline surveillance for 2008-09 (USA and Europe) documented low MIC(50/90) values for S. aureus isolates at 0.5/1 and 0.25/1 mg/L, respectively. More importantly, ceftaroline MIC(90) results for S. pneumoniae (0.12 mg/L), H. influenzae (0.015 mg/L) and M. catarrhalis (0.12 mg/L) were very low, all MICs being ≤0.5 mg/L. Ceftaroline exhibited promising high potency and wide coverage against Gram-positive and -negative pathogens known to cause CAP, especially isolates of MDR pneumococci and methicillin-resistant S. aureus.


Air and Bone Conduction Click and Tone-Burst Auditory Brainstem Thresholds Using Kalman Adaptive Processing in Nonsedated Normal-Hearing Infants.
To study normative thresholds and latencies for click and tone-burst auditory brainstem response (TB-ABR) for air and bone conduction in normal infants and those discharged from neonatal intensive care units, who passed newborn hearing screening and follow-up distortion product otoacoustic emission. An evoked potential system (Vivosonic Integrity) that incorporates Bluetooth electrical isolation and Kalman-weighted adaptive processing to improve signal to noise ratios was employed for this study. Results were compared with other published data. One hundred forty-five infants who passed two-stage hearing screening with transient-evoked otoacoustic emission or automated auditory brainstem response were assessed with clicks at 70 dB nHL and threshold TB-ABR. Tone bursts at frequencies between 500 and 4000 Hz were used for air and bone conduction auditory brainstem response testing using a specified staircase threshold search to establish threshold levels and wave V peak latencies. Median air conduction hearing thresholds using TB-ABR ranged from 0 to 20 dB nHL, depending on stimulus frequency. Median bone conduction thresholds were 10 dB nHL across all frequencies, and median air-bone gaps were 0 dB across all frequencies. There was no significant threshold difference between left and right ears and no significant relationship between thresholds and hearing loss risk factors, ethnicity, or gender. Older age was related to decreased latency for air conduction. Compared with previous studies, mean air conduction thresholds were found at slightly lower (better) levels, while bone conduction levels were better at 2000 Hz and higher at 500 Hz. Latency values were longer at 500 Hz than previous studies using other instrumentation. Sleep state did not affect air or bone conduction thresholds. This study demonstrated slightly better wave V thresholds for air conduction than previous infant studies. The differences found in the present study, while statistically significant, were within the test step size of 10 dB. This suggests that threshold responses obtained using the Kalman weighting software were within the range of other published studies using traditional signal averaging, given step-size limitations. Thresholds were not adversely affected by variable sleep states.


Habitat associations of two entomopathogenic nematodes: a quantitative study using real-time quantitative polymerase chain reactions.
1. Despite nematodes being the most abundant animals on earth, very few animal ecologists study them, probably because of the difficulties of identifying them to species by morphological methods. 2. A group of nematodes that are important both ecologically and economically is the entomopathogenic nematodes, which play a key role in regulating soil food webs and are sold throughout the world as biological insecticides, yet for which very little is known of their population ecology. 3. A novel detection and quantification method was developed for soil nematodes using real-time polymerase chain reaction (PCR), and the technique was used to estimate numbers of two closely related species of entomopathogenic nematodes, Steinernema kraussei and S. affine in 50 soil samples from 10 sites in Scotland representing two distinct habitats (woodland and grassland). 4. There was a high degree of correlation between our molecular and traditional morphological estimates of population size and our data clearly showed that Steinernema affine occurred only in grassland areas, whereas S. kraussei was found in grassland and woodland samples to a similar degree. 5. Real-time PCR offers a rapid and accurate method of detecting individual nematode species from soil samples without the need for a specialist taxonomist, and has much potential for use in studies of nematode population ecology.


Systemic lupus erythematosus children in Egypt: Homeland spectrum amid the global situation.
This study aims to present the manifestations of juvenile systemic lupus erythematosus (JSLE) across Egypt, to focus on age at onset and gender-driven influence on disease characteristics, and to compare findings to other countries. The study included 404 Egyptian children with systemic lupus erythematosus (SLE) presenting to one of the specialized rheumatology centers corresponding to 13 major governorates. Juvenile cases age was ≤ 16°years at the time of recruitment. The SLE Disease Activity Index (SLEDAI) and damage index (DI) were assessed. The mean age was 13.2 ± 2.4°years; 355 females and 49 males (7.2:1), and the disease duration was 2.3 ± 1.6 years, while age at disease onset was 11.1 ± 2.5°years. Their SLEDAI was 13.5 ± 12.3, and DI, 0.36 ± 0.78. The overall estimated prevalence of childhood-SLE patients in the recruited cohort in Egypt was 1/100,000 population (0.24/100000 males and 1.8/100000 females). 7.4% developed pre-pubertal SLE (≤ 7 years); 73.3%, peri-pubertal; and 19.3% during early adolescence. The differences according to age group were equal for gender and clinical manifestations except skin lesions present in 59.3% of pre-pubertal onset, 74.6% of peri-pubertal, and 84.2% of adolescents (p = 0.029), and renal involvement in 73.8% of peripubertal, 62.1% of pre-pubertal and 58.9% of adolescents (p = 0.03). Laboratory investigations, SLEDAI, and DI were similar among age categories. Lupus nephritis was more common in Egypt compared to JSLE from other countries. Our large multicenter study identified that female gender influenced disease characteristics with more frequent skin involvement. Skin lesions were significantly higher in adolescents, while renal involvement in peri-pubertal children.


Five-Year Hearing Outcomes in Bilateral Simultaneously Cochlear-Implanted Adult Patients.
To report the speech performance and sound localization in adult patients 5 years after bilateral simultaneous cochlear implantation and to evaluate the change in speech scores between 1 and 5 years. In this prospective multicenter study, 26 patients were evaluated 5 years after implantation using long straight electrode arrays (MED-EL Combi 40+, standard electrode array, 31 mm). Speech perception was measured using disyllabic words in quiet and noise, with the speech coming from the front and a cocktail party background noise coming from 5 loudspeakers. Speech localization measurements were performed in noise under the same test conditions. These results were compared to those obtained at 1 year reported in a previous study. Five years after implantation, an improvement in speech performance scores compared to 1 year after implantation was found for the poorer ear both in quiet and in noise (+12.1 ± 2.6%, p < 0.001). The lower the speech score of the poorer ear at 1 year, the greater the improvement at 5 years, both in quiet (r = -0.62) and at a signal-to-noise ratio of +15 dB (r = -0.58). The sound localization on the horizontal plane in noise provided by bilateral implantation was better than the unilateral one and remained stable after the results observed at 1 year. In adult patients simultaneously and bilaterally implanted, the poorest speech scores improved between 1 and 5 years after implantation. These findings are an additional element to recommend bilateral implantation in adult patients. The use of both cochlear implants and speech training sessions for patients with poor performance should continue in the period after 1 year following implantation, since the speech scores will improve over time.


Multimeric forms of rat liver glyoxalase II.
Rat liver glyoxalase II has been purified to homogeneity by a rapid, two-step procedure involving affinity chromatography on S-carbobenzoxyglutathione coupled to Sepharose 4B. The purified enzyme gives a major band corresponding to 30,000 daltons (monomer) and a minor band corresponding to 120,000 daltons (tetramer) with sodium dodecylsulfate polyacrylamide gel electrophoresis. Evidence is given for the interconversion of the monomer and tetramer forms of glyoxalase II via the dimer and the trimer.


Eruption of squamous cell carcinomas after beginning nilotinib therapy.
Chronic myelogenous leukemia (CML) is characterized by a reciprocal translocation between the long arms of chromosomes 9 and 22 leading to the formation of a constitutively active tyrosine kinase. Tyrosine kinase inhibitors (TKIs) are the treatment of choice for patients diagnosed with CML and have many associated side effects including the rarely-reported eruption of squamous cell carcinomas (SCCs). Herein, we report a patient with CML who presented with sudden onset of multiple scaly lesions on his legs and trunk after beginning treatment with nilotinib, a novel TKI. Six biopsies were performed at his initial presentation and four of these lesions were confirmed to be keratoacanthoma-type SCCs. One month later, the patient reported the development of multiple new similar lesions on his legs, arms, and face. Four more biopsies were performed revealing keratoacanthoma-type and well-differentiated SCCs. Certain tyrosine kinase inhibitors such as sorafenib and quizartinib have been reported to cause eruptive keratoacanthoma (KA)-type SCCs as seen in our patient. However, there is only one other report in the literature of nilotinib promoting the development of SCCs or KAs. Physicians should be aware of this potential adverse effect and patients taking nilotinib should be closely monitored by a dermatologist.


Assembly and suppression of endogenous Kv1.3 channels in human T cells.
The predominant K+ channel in human T lymphocytes is Kv1.3, which inactivates by a C-type mechanism. To study assembly of these tetrameric channels in Jurkat, a human T-lymphocyte cell line, we have characterized the formation of heterotetrameric channels between endogenous wild-type (WT) Kv1.3 subunits and heterologously expressed mutant (A413V) Kv1.3 subunits. We use a kinetic analysis of C-type inactivation of currents produced by homotetrameric channels and heterotetrameric channels to determine the distribution of channels with different subunit stoichiometries. The distributions are well-described by either a binomial distribution or a binomial distribution plus a fraction of WT homotetramers, indicating that subunit assembly is a random process and that tetramers expressed in the plasma membrane do not dissociate and reassemble. Additionally, endogenous Kv1.3 current is suppressed by a heterologously expressed truncated Kv1.3 that contains the amino terminus and the first two transmembrane segments. The time course for suppression, which is maximal at 48 h after transfection, overlaps with the time interval for heterotetramer formation between heterologously expressed A413V and endogenous WT channels. Our findings suggest that diversity of K+ channel subtypes in a cell is regulated not by spatial segregation of monomeric pools, but rather by the degree of temporal overlap and the kinetics of subunit expression.


The 2013 WHO guidelines for antiretroviral therapy: evidence-based recommendations to face new epidemic realities.
The review summarizes the key new recommendations of the WHO 2013 guidelines for antiretroviral therapy and describes the potential impact of these recommendations on the HIV epidemic. The 2013 WHO guidelines recommend earlier initiation of antiretroviral therapy (ART) at CD4 500 cells/μl or less for all adults and children above 5 years. Further recommendations include initiation of ART irrespective of CD4 cell count or clinical stage for people co-infected with active tuberculosis disease or hepatitis B virus with severe liver disease, pregnant women, people in serodiscordant partnerships, and children under 5 years of age. The ART regimen comprising a once daily fixed-dose combination of tenofovir + lamivudine (or emtricitabine) + efavirenz is recommended as the preferred first-line therapy. Several approaches are also recommended to reach more people and increase health service capacity, including community and self-testing, and task shifting, decentralization, and integration of ART care. If fully implemented, the 2013 WHO ART guidelines could avert at least an additional 3 million deaths and prevent close to an additional 3.5 million new infections between 2012 and 2025 in low- and middle-income countries, compared with previous treatment guidelines.


Combined inhibition of c-Src and epidermal growth factor receptor abrogates growth and invasion of head and neck squamous cell carcinoma.
Increased expression and/or activation of epidermal growth factor receptor (EGFR) is associated with tumor progression and poor prognosis in many cancers, including head and neck squamous cell carcinoma (HNSCC). Src family kinases, including c-Src, mediate a variety of intracellular or extracellular signals that contribute to tumor formation and progression. This study was undertaken to elucidate the role of c-Src in the growth and invasion of HNSCC and to determine the effects of combined targeting of EGFR and Src kinases in HNSCC cell lines. HNSCC cells were engineered to stably express a dominant-active form of c-Src and investigated in cell growth and invasion assays. The biochemical effects of combined treatment with the Src inhibitor AZD0530, a potent, orally active Src inhibitor with Bcr/Abl activity, and the EGFR kinase inhibitor gefitinib were examined, as well as the consequences of dual Src/EGFR targeting on the growth and invasion of a panel of HNSCC cell lines. HNSCC cells expressing dominant-active c-Src showed increased growth and invasion compared with vector-transfected controls. Combined treatment with AZD0530 and gefitinib resulted in greater inhibition of HNSCC cell growth and invasion compared with either agent alone. These results suggest that increased expression and activation of c-Src promotes HNSCC progression where combined targeting of EGFR and c-Src may be an efficacious treatment approach.


Mental models, metaphors and their use in the education of nurses.
A great deal of nurses' confidence in the use of information technology (IT) depends both on the way computers are introduced to students in the college and how such education is continued and applied when they are practitioners. It is therefore vital that teachers of IT assist nurses to discover ways of learning to utilize and apply computers within their workplace with whatever methods are available. One method which has been introduced with success in other fields is the use of mental models and metaphors. Mental models and metaphors enable individuals to learn by building on past learning. Concepts and ideas which have already been internalized from past experience can be transferred and adapted for usage in a new learning situation with computers and technology. This article explores the use of mental models and metaphors for the technological education of nurses. The concepts themselves will be examined, followed by suggestions for possible applications specifically in the field of nursing and health care. Finally the role of the teacher in enabling improved learning as a result of these techniques will be addressed.


The Role of Architectural and Learning Constraints in Neural Network Models: A Case Study on Visual Space Coding.
The recent "deep learning revolution" in artificial neural networks had strong impact and widespread deployment for engineering applications, but the use of deep learning for neurocomputational modeling has been so far limited. In this article we argue that unsupervised deep learning represents an important step forward for improving neurocomputational models of perception and cognition, because it emphasizes the role of generative learning as opposed to discriminative (supervised) learning. As a case study, we present a series of simulations investigating the emergence of neural coding of visual space for sensorimotor transformations. We compare different network architectures commonly used as building blocks for unsupervised deep learning by systematically testing the type of receptive fields and gain modulation developed by the hidden neurons. In particular, we compare Restricted Boltzmann Machines (RBMs), which are stochastic, generative networks with bidirectional connections trained using contrastive divergence, with autoencoders, which are deterministic networks trained using error backpropagation. For both learning architectures we also explore the role of sparse coding, which has been identified as a fundamental principle of neural computation. The unsupervised models are then compared with supervised, feed-forward networks that learn an explicit mapping between different spatial reference frames. Our simulations show that both architectural and learning constraints strongly influenced the emergent coding of visual space in terms of distribution of tuning functions at the level of single neurons. Unsupervised models, and particularly RBMs, were found to more closely adhere to neurophysiological data from single-cell recordings in the primate parietal cortex. These results provide new insights into how basic properties of artificial neural networks might be relevant for modeling neural information processing in biological systems.


Bovine Lactoferrin Prevents Influenza A Virus Infection by Interfering with the Fusogenic Function of Viral Hemagglutinin.
Bovine lactoferrin (bLf) is an iron-binding glycoprotein folded in two symmetric globular lobes (N- and C-lobes) with potent antimicrobial and immunomodulatory activities. Recently, we have shown that bLf, and in particular its C-lobe, interacts with influenza A virus hemagglutinin and prevents infection by different H1 and H3 viral subtypes. Influenza virus hemagglutinin (HA), and in particular its highly conserved fusion peptide involved in the low-pH-mediated fusion process, plays a significant role in the early steps of viral infection and represents an attractive target for the development of anti-influenza drugs. In the present research, we further investigated the influence of low pH on the interactions between bLf and influenza A H1N1 virus by different techniques, such as enzyme-linked immunosorbent assay, electron microscopy, hemolysis inhibition assay, and time course assay. Our results demonstrate that lactoferrin interaction with influenza hemagglutinin at low pH induces alterations that stabilize the conformation of the hemagglutinin, resulting in the inhibition of the fusion peptide activity. Taken together, our data allowed to better characterize the HA-specific inhibiting activity of bLf and to confirm HA as a good target for drug development.


The organization of the tadpole and adult alpha globin genes of Xenopus laevis.
Adult erythrocytes of X. laevis contain six electrophoretically resolvable globin polypeptides while tadpole erythrocytes contain four polypeptides, none of which comigrates with an adult protein. We show that three of the adult proteins are alpha globin polypeptides (alpha 1, alpha 2, alpha 3) and three are beta globin polypeptides (beta 1, beta 2, beta 3). We find that a tadpole alpha globin gene (alpha T1) is linked to the major adult locus in the sequence 5'-alpha T1-alpha 1-beta 1-3' with 5.2 kb separating alpha T1 from alpha 1. Another tadpole alpha globin gene (alpha T2) is linked to the minor adult locus in the sequence 5'-alpha T2-alpha 2-beta 2-3' with 10.7 kb separating alpha T2 from alpha 2. These linkage relationships are consistent with the major and minor loci having arisen by tetraploidization but the different separation of larval and adult globin genes at the two loci indicates the occurrence of some additional chromosomal rearrangement. Two alternative models are presented.


Clinical remission in severe asthma with biologic therapy: an analysis from the UK Severe Asthma Registry.
Novel biologic therapies have revolutionised the management of severe asthma with more ambitious treatment aims. Here we analyse the definition of clinical remission as a suggested treatment goal and consider the characteristics associated with clinical remission in a large, real-world severe asthma cohort. This was a retrospective analysis of severe asthma patients registered in the UK Severe Asthma Registry (UKSAR) who met strict national access criteria for biologics. Patients had a pre-biologics baseline assessment and annual review. The primary definition of clinical remission applied included Asthma Control Questionnaire (ACQ)-5 <1.5 and no oral corticosteroids for disease control and forced expiratory volume in 1 s above lower limit of normal or no more than 100 mL less than baseline. 18.3% of patients achieved the primary definition of remission. The adjusted odds of remission on biologic therapy were 7.44 (95% CI 1.73-31.95)-fold higher in patients with type 2 (T2)-high biomarkers. The adjusted odds of remission were lower in patients who were female (OR 0.61, 95% CI 0.45-0.93), obese (OR 0.49, 95% CI 0.24-0.65) or had ACQ-5 ≥1.5 (OR 0.19, 95% CI 0.12-0.31) pre-biologic therapy. The likelihood of remission reduced by 14% (95% CI 0.76-0.97) for every 10-year increase in disease duration. 12-21% of the cohort attained clinical remission depending on the definition applied; most of those who did not achieve remission failed to meet multiple criteria. 18.3% of patients achieved the primary definition of clinical remission. Remission was more likely in T2-high biomarker patients with shorter duration of disease and less comorbidity. Further research on the optimum time to commence biologics in severe asthma is required.


Acute change in the cyclic AMP content of rat mammary acini in vitro. Influence of physiological and pharmacological agents.
The cyclic AMP content of acini, freshly prepared from mammary tissue of lactating rats, was measured during incubation in vitro. Neither adrenergic agonists nor cyclic AMP phosphodiesterase inhibitors alone caused a change of more than 2-fold in the basal cyclic AMP content of acini. Together, however, these agents provoked increases of around 20-fold in acini cyclic AMP content. Forskolin caused similar effects. The relative potency of adrenergic agonists in increasing cyclic AMP in acini, together with the ability of selective antagonists to oppose such rises, indicated that beta 2-adrenergic receptors were involved in mediating the effects. Receptor-binding experiments using [3H]dihydroalprenolol and selective beta-antagonists confirmed the predominant presence of beta 2-adrenergic receptors on acini membranes and on membranes prepared from purified mammary secretory epithelial cells. These results elucidate some previous findings [Robson, Clegg & Zammit (1984) Biochem. J. 217, 743-749; Williamson, Munday, Jones, Roberts & Ramsey (1983) Adv. Enzyme Regul. 21, 135-145], questioning the role of cyclic AMP in the regulation of lipogenesis in mammary acini.


Characteristics of the atrial repolarization phase of the ECG in paroxysmal atrial fibrillation patients and controls.
The aim of this study is to characterize the observable segment of the atrial repolarization (Ta wave) of the standard ECG during sinus rhythm in paroxysmal atrial fibrillation (PAF) patients and controls. Ta and P waves were measured from signal-averaged recordings of a standard 12-lead ECG in 40 patients, 20 with PAF, but in SR at the time of recording, and 20 healthy controls. Wave amplitudes and morphologies were measured. There were no significant differences in Ta amplitude between the PAF patients and controls. A subgroup analysis of patients on and off anti-arrhythmic drugs also showed no significant differences in Ta amplitudes. For both groups Ta wave had opposite polarity to the monophasic P wave. Biphasic P waves had Ta polarity opposite to the initial phase of the P wave. Ta wave amplitudes were largest in leads II (mean ± SD, –25 ± 16 μV), V2 (–22 ± 10 μV), V3 (–21 ± 10 μV) and V4 (–20 ± 8 μV). A significant correlation was found between Ta and P wave amplitudes, leads recording larger P waves also had larger Ta waves (PAF group: r = 0.15 (P = 0.02) PAF vs r = 0.33 (P = 0.002) HC). No differences in the amplitude of the observable section of the atrial repolarization phase of the ECG could be observed between patients with PAF and controls. Ta wave correlates with the corresponding P wave in both amplitude and polarity.


Gambling in COVID-19 Lockdown in the UK: Depression, Stress, and Anxiety.
To combat the spread of COVID-19, the UK Government implemented a range of "lockdown" measures. Lockdown has necessarily changed the gambling habits of gamblers in the UK, and the impact of these measures on the mental health of gamblers is unknown. To understand the impact of lockdown on gamblers, in April 2020, after ~6 weeks of lockdown, participants (N = 1,028, 72% female) completed an online questionnaire. Gambling engagement data was collected for pre-lockdown via the Brief Problem Gambling Screen (BPGS) allowing participants to be classified as Non-Gamblers (NG), Non-Problem Gamblers (NPG) or Potential Problem Gamblers (PPG). The Depression, Stress, and Anxiety Scale (DASS21) was used to measure depression, stress, and anxiety scores both pre- and during-lockdown. Results indicate that depression, stress and anxiety has increased across the whole sample. Participants classified in the PPG group reported higher scores on each sub scale at both baseline and during lockdown. Increases were observed on each DASS21 subscale, for each gambler group, however despite variable significance and effect sizes, the magnitude of increases did not differ between groups. Lockdown has had a significant impact on mental health of participants; whilst depression stress and anxiety remain highest in potential problem gamblers, pre-lockdown gambler status did not affect changes in DASS21 scores.